中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2009年
3期
233-239
,共7页
刘琼%赵世华%陆敏杰%蒋世良%闫朝武%张岩%孟亮%唐跃%孟宪敏%魏英杰%王琳琳%戴鸿君%徐坚
劉瓊%趙世華%陸敏傑%蔣世良%閆朝武%張巖%孟亮%唐躍%孟憲敏%魏英傑%王琳琳%戴鴻君%徐堅
류경%조세화%륙민걸%장세량%염조무%장암%맹량%당약%맹헌민%위영걸%왕림림%대홍군%서견
心肌梗死%磁共振成像%干细胞移植
心肌梗死%磁共振成像%榦細胞移植
심기경사%자공진성상%간세포이식
Myocardial infarction%Magnetic resonance imaging%Stem cell transplantation
目的 通过观察血管内皮生长因子(VEGF)缓释明胶微球与自体骨髓间充质干细胞(MSC)联合移植于猪心肌梗死模型梗死周边区3周后MSC的生存情况,结合磁共振成像(MRI),阐明改善局部微环境对自体MSC移植于缺血心肌后转归的影响,并直观评价MRI对移植MSC在体示踪及半定量分析的可靠性.方法 以乳化冷凝法制备VEGF缓释明胶微球并评价其缓释性能.成年中华小型猪12头,抽取髂骨骨髓并制备自体MSC,以顺磁性氧化铁颗粒(SPIO)和4',6-二脒基-2-苯基吲哚(4',6-Diamidino-2-phenylindole dihydrochloride,DAPI)标记细胞.按照随机化表将动物分为MSC移植组及MSC-VEGF缓释微球联合移植组.开胸结扎冠状动脉左前降支制备小型猪心肌梗死模型后第14天,直视下经心外膜于MSC组动物心肌梗死周边区注射MSC(9×107),于MSC-VEGF缓释微球组注射MSC(9×107)及VEGF缓释明胶微球(含9 μg人重组VEGF165).细胞和(或)微球移植后24 h及21 d,磁共振FGRE序列T2*像检测干细胞移植点低信号区的面积及信号强度.病理组织学检测细胞移植区域心肌组织毛细血管密度及干细胞存活情况.结果 明胶微球平均粒径为(104.0±22.6)μm,体内、外缓释VEGF均可达到10 d以上.小型猪心肌梗死模型建立后第14天,梗死区面积为33.6%±8.9%.细胞移植后24 h,MSC注射点于MRI显示为边界清晰的卵圆形T2*低信号区,细胞移植3周后,两组注射点T2*低信号区面积均减小(P<0.05),其与正常心肌组织间的信号对比度均减低(P<0.05).MSC-VEGF缓释微球组移植点毛细血管密度高于MSC组[(15.2±5.4)个/高倍视野比(10.2±5.0)个/高倍视野,t=2.43,P<0.05].MSC-VEGF缓释微球组MSC注射点移植细胞数多于MSC组[(354±83)个/高倍视野比(278±97)个/高倍视野,t=3.14,P<0.05],而MSC凋亡率则小于MSC-VEGF缓释微球组[(6.4±4.1)%比(11.9±4.8)%,t=2.97,P<0.05].结论 VEGF缓释明胶微球可以改善移植于缺血心肌区3周后MSC的生存情况.移植干细胞所生存的微环境是影响其转归的重要因素.MRI对于移植干细胞位置的在体示踪是可靠的,但不能作为对移植细胞定量或半定量分析的依据.
目的 通過觀察血管內皮生長因子(VEGF)緩釋明膠微毬與自體骨髓間充質榦細胞(MSC)聯閤移植于豬心肌梗死模型梗死週邊區3週後MSC的生存情況,結閤磁共振成像(MRI),闡明改善跼部微環境對自體MSC移植于缺血心肌後轉歸的影響,併直觀評價MRI對移植MSC在體示蹤及半定量分析的可靠性.方法 以乳化冷凝法製備VEGF緩釋明膠微毬併評價其緩釋性能.成年中華小型豬12頭,抽取髂骨骨髓併製備自體MSC,以順磁性氧化鐵顆粒(SPIO)和4',6-二脒基-2-苯基吲哚(4',6-Diamidino-2-phenylindole dihydrochloride,DAPI)標記細胞.按照隨機化錶將動物分為MSC移植組及MSC-VEGF緩釋微毬聯閤移植組.開胸結扎冠狀動脈左前降支製備小型豬心肌梗死模型後第14天,直視下經心外膜于MSC組動物心肌梗死週邊區註射MSC(9×107),于MSC-VEGF緩釋微毬組註射MSC(9×107)及VEGF緩釋明膠微毬(含9 μg人重組VEGF165).細胞和(或)微毬移植後24 h及21 d,磁共振FGRE序列T2*像檢測榦細胞移植點低信號區的麵積及信號彊度.病理組織學檢測細胞移植區域心肌組織毛細血管密度及榦細胞存活情況.結果 明膠微毬平均粒徑為(104.0±22.6)μm,體內、外緩釋VEGF均可達到10 d以上.小型豬心肌梗死模型建立後第14天,梗死區麵積為33.6%±8.9%.細胞移植後24 h,MSC註射點于MRI顯示為邊界清晰的卵圓形T2*低信號區,細胞移植3週後,兩組註射點T2*低信號區麵積均減小(P<0.05),其與正常心肌組織間的信號對比度均減低(P<0.05).MSC-VEGF緩釋微毬組移植點毛細血管密度高于MSC組[(15.2±5.4)箇/高倍視野比(10.2±5.0)箇/高倍視野,t=2.43,P<0.05].MSC-VEGF緩釋微毬組MSC註射點移植細胞數多于MSC組[(354±83)箇/高倍視野比(278±97)箇/高倍視野,t=3.14,P<0.05],而MSC凋亡率則小于MSC-VEGF緩釋微毬組[(6.4±4.1)%比(11.9±4.8)%,t=2.97,P<0.05].結論 VEGF緩釋明膠微毬可以改善移植于缺血心肌區3週後MSC的生存情況.移植榦細胞所生存的微環境是影響其轉歸的重要因素.MRI對于移植榦細胞位置的在體示蹤是可靠的,但不能作為對移植細胞定量或半定量分析的依據.
목적 통과관찰혈관내피생장인자(VEGF)완석명효미구여자체골수간충질간세포(MSC)연합이식우저심기경사모형경사주변구3주후MSC적생존정황,결합자공진성상(MRI),천명개선국부미배경대자체MSC이식우결혈심기후전귀적영향,병직관평개MRI대이식MSC재체시종급반정량분석적가고성.방법 이유화냉응법제비VEGF완석명효미구병평개기완석성능.성년중화소형저12두,추취가골골수병제비자체MSC,이순자성양화철과립(SPIO)화4',6-이미기-2-분기신타(4',6-Diamidino-2-phenylindole dihydrochloride,DAPI)표기세포.안조수궤화표장동물분위MSC이식조급MSC-VEGF완석미구연합이식조.개흉결찰관상동맥좌전강지제비소형저심기경사모형후제14천,직시하경심외막우MSC조동물심기경사주변구주사MSC(9×107),우MSC-VEGF완석미구조주사MSC(9×107)급VEGF완석명효미구(함9 μg인중조VEGF165).세포화(혹)미구이식후24 h급21 d,자공진FGRE서렬T2*상검측간세포이식점저신호구적면적급신호강도.병리조직학검측세포이식구역심기조직모세혈관밀도급간세포존활정황.결과 명효미구평균립경위(104.0±22.6)μm,체내、외완석VEGF균가체도10 d이상.소형저심기경사모형건립후제14천,경사구면적위33.6%±8.9%.세포이식후24 h,MSC주사점우MRI현시위변계청석적란원형T2*저신호구,세포이식3주후,량조주사점T2*저신호구면적균감소(P<0.05),기여정상심기조직간적신호대비도균감저(P<0.05).MSC-VEGF완석미구조이식점모세혈관밀도고우MSC조[(15.2±5.4)개/고배시야비(10.2±5.0)개/고배시야,t=2.43,P<0.05].MSC-VEGF완석미구조MSC주사점이식세포수다우MSC조[(354±83)개/고배시야비(278±97)개/고배시야,t=3.14,P<0.05],이MSC조망솔칙소우MSC-VEGF완석미구조[(6.4±4.1)%비(11.9±4.8)%,t=2.97,P<0.05].결론 VEGF완석명효미구가이개선이식우결혈심기구3주후MSC적생존정황.이식간세포소생존적미배경시영향기전귀적중요인소.MRI대우이식간세포위치적재체시종시가고적,단불능작위대이식세포정량혹반정량분석적의거.
Objective To investigate the efficacy of transplantation of mesenchymal stem cells(MSC) with gelatin micrespheres containing vascular endothelial growth factor in ischemie regions in infracted swine hearts. Methods Twelve Chinese mini swines with infarction were randomized to receive autogenetic MSC injection to the peri-infarction area of left ventricular wall (MSC group, n=6) or MSC transplantation with gelatin hydrogel microspheres incorporating vascular endothelial growth factor (VEGF-MSC group, n=6). Three weeks later, left ventricular function was assessed by magnetic resonance imaging (MRI). The contrast of the MSC hypointense lesion was determined using the difference in signal intensity between the hypointenso and normal myocardium divided by signal intensity of the normal region. Myocardial capillary density, the number of DAPI positive MSC and the apoptotic MSC were also determined. Results The diameter of the microspheres averaged (104.0±22.6) μm. At 24 hours after transplantation, MSC were identified by MRI as large intramyocardial signal voids at injection sites which persisted up to 3 weeks. There was no significant difference in the contrast of the lesions and in the size of the lesions at 24 hours between two groups. At 3 weeks after injection, the size of the lesions and the contrast of the lesion were decreased (P<0.05) in both groups. The capillary density of the injection site was significantly more in the MSC-VEGF microsphere group than that in MSC group [ (15.2±5.4)/HPF vs. (10.2±5.0)/HPF, t=2.43, P<0.05], and there were more dense DAPI labeled MSC per high power fields in injection sites of MSC-VEGF mieresphere group than that in MSC group [(354±83)/HPF vs. (278±97)/HPF, t=3.14, P<0.05]. Moreover, the apoptosis rate of MSCs of MSCs-VEGF mieresphere group was less than that of MSCgroup [(6.4±4.1)% vs. (11.9±4.8)%, t=2.97, P<0.05]. Conclusions MSC transplantation with gelatin hydrogel microspheres incorporating VEGF enhanced the efficacy of MSC in this swine model of myocardial infarction. MRI tracking of MSC is feasible and represents a preferred method for studying the engraftment of MSCs in infracted tissue.
